ArticleGenome biology2018
Manipulating plant RNA-silencing pathways to improve the gene editing efficiency of CRISPR/Cas9 systems.
Article in Genome biology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 53 citations in OpenAlex.
- Advancements in CRISPR-Based Therapies for Genetic Modulation in Neurodegenerative Disorders.Current gene therapy · 2025Review
- Modern Plant Breeding Techniques in Crop Improvement and Genetic Diversity: From Molecular Markers and Gene Editing to Artificial Intelligence-A Critical Review.Plants (Basel, Switzerland) · 2024Review
- Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding.Plants (Basel, Switzerland) · 2023Review
- Establishment of targeted mutagenesis in soybean protoplasts using CRISPR/Cas9 RNP delivery via electro-transfection.Frontiers in plant science · 2023Article
- Review
- Genome Editing Targets for Improving Nutrient Use Efficiency and Nutrient Stress Adaptation.Frontiers in genetics · 2022Review
- Expanding the target range of base editing in plants without loss of efficiency by blocking RNA-silencing.Plant biotechnology journal · 2021Article
- Advances in application of genome editing in tomato and recent development of genome editing technology.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2021Review
- Novel CRISPR/Cas applications in plants: from prime editing to chromosome engineering.Transgenic research · 2021Review
- CRISPR-based targeting of DNA methylation inProceedings of the National Academy of Sciences of the United States of America · 2021Article
- An Efficient Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/CRISPR-Associated Protein 9 Mutagenesis System for Oil Palm (Frontiers in plant science · 2021Article
- Targets and Mechanisms of Geminivirus Silencing Suppressor Protein AC2.Frontiers in microbiology · 2021Review
- Non-GM Genome Editing Approaches in Crops.Frontiers in genome editing · 2021Review
- Review
- An RNAi suppressor activates in planta virus-mediated gene editing.Functional & integrative genomics · 2020Article
- RNAi-mediated control of CRISPR functions.Theranostics · 2020Article
- Protein expression and gene editing in monocots using foxtail mosaic virus vectors.Plant direct · 2019Article
- Article
- 3D-printing enabled micro-assembly of a microfluidic electroporation system for 3D tissue engineering.Lab on a chip · 2019Article
- Cas9-PF, an early flowering and visual selection marker system, enhances the frequency of editing event occurrence and expedites the isolation of genome-edited and transgene-free plants.Plant biotechnology journal · 2019Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 3 countries.
Funding
Abstract
backgroundThe CRISPR/Cas9 system, composed of a single-guide RNA for target recognition and a Cas9 protein for DNA cleavage, has the potential to revolutionize agriculture as well as medicine. Even though extensive work has been done to improve the gene editing activity of CRISPR/Cas9, little is known about the regulation of this bacterial system in eukaryotic host cells, especially at the post-transcriptional level.
resultsHere, we evaluate the expression levels of the two CRISPR/Cas9 components and the gene editing efficiency in a set of Arabidopsis mutants involved in RNA silencing. We find that mutants defective in the post-transcriptional gene-silencing pathway display significantly higher Cas9 and sgRNA transcript levels, resulting in higher mutagenesis frequencies than wild-type controls. Accordingly, silencing of AGO1 by introduction of an AGO1-RNAi cassette into the CRISPR/Cas9 vector provides an increase in gene editing efficiency. Co-expression of the viral suppressor p19 from the tomato bushy stunt virus to suppress the plant RNA-silencing pathway shows a strong correlation between the severity of the phenotypic effects caused by p19 and the gene editing efficiency of the CRISPR/Cas9 system for two different target genes, AP1 and TT4.
conclusionsThis system has useful practical applications in facilitating the detection of CRISPR/Cas9-induced mutations in T1 plants as well as the identification of transgene-free T2 plants by simple visual observation of the symptom severity caused by p19. Our study shows that CRISPR/Cas9 gene editing efficiency can be improved by reducing RNA silencing in plants.
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